Project Summary/Abstract TIC10 (ONC201) is a first-in-class antitumor agent and small molecule inducer of the TRAIL gene which was identified in a high-throughput small molecule library screen in the lab of our collaborator, Dr. Wafik El-Deiry at Penn State University and has been shown to have preclinical efficacy in several difficult-to-treat disease settings that include unmet clinical needs such as glioblastoma and triple-negative breast cancer. The motivation for developing the small molecule TIC10 and analogs is to overcome efficacy-limiting properties of recombinant TRAIL and to optimize advantages TIC10 that have been demonstrated (Allen et al, 2013), including: (i) longer half-life, (ii) prolonged elevation of serum TRAIL, (iii) a bystander effect through normal cell TRAIL production, (iv) stimulation of TRAIL and death receptor expression, and other properties that are only obtained by a small molecule, such as (v) stability, (vi) cost, (vii) oral activity, and (viii) ability to cross the blood-brain barrier. Based on promising efficacy and mechanistic studies with TIC10, efforts have begun to pursue preclinical development of the parent TIC10 compound through our collaborating development company, Oncoceutics Inc. toward a proof-of-concept trial in humans. The project proposed here is intended to identify more potent analogs of TIC10 that maintain the efficacy and safety profile of TIC10 but are novel and suitable for commercial development. While the TIC10 molecule was originally synthesized in 1971, TIC10 was the single example of this ring system available for evaluation in our assays and except for TIC10, this structural class has not been previously studied for biological activity. We propose to synthesize TIC10 analogs to identify a more potent lead compound and to develop structure-activity relationship (SAR) data that would facilitate optimization of activity while generating novel, patentable analogs. The project proposed here is intended to leverage our chemistry and biology experience to synthesize and characterize analogs and to explore SAR (Aim1) and to optimize activity in cellular and in vivo models and to enhance pharmaceutical properties (Aim2). The exploratory work in this phase I application is also planned to generate proprietary variants and novel composition of matter intellectual property that would establish a pathway toward drug lead optimization, future clinical trials and commercialization for the treatment of difficult cancers. Together, these studies are aimed to increase our basic and translational understanding of TRAIL- inducing small molecules and to uncover a superior lead compound that would advance this novel class of compounds to the clinic to offer therapeutic benefit to cancer patients without other treatment options.